Zebrafish Mate7 belongs to solute carrier protein superfamily and specifically to subfamily of multidrug and toxin extruders. It is co-orthologous to mammalian Mates, and is ubiquitously expressed in zebrafish tissues with the highest expression in kidney. It has been shown to interact with both endogenous (steroid hormones) and xenobiotic compounds (pharmaceuticals), implying a role in efflux of toxic compounds. The objective of our study was to analyse interaction of environmental contaminants with zebrafish Mate7 using a newly developed high throughput screening (HTS) Mate7 assay. A full-length zebrafish mate7 sequence was obtained from zebrafish cDNA originating from male kidney, and a stable expression of Mate7 in genetically engineered HEK293 Flp-In cells was achieved. Stable Mate7 transfectants were then used for development and optimization of a new HTS cellular uptake protocol, with DAPI and ASP + as model fluorescent substrates. The developed assay was used for identifying zebrafish Mate 7 interactors and discerning the type of interaction. A series of 89 diverse environmental contaminants, including industrial chemicals, pesticides, and pharmaceuticals, was tested and highly effective Mate7 interactors were identified in all of the aforementioned groups. Some of the inhibitors identified could be of environmental concern because they may potentially impair Mate7 efflux function, lowering the fish defence capacity against environmental contaminants, or interfering with transport of yet unidentified physiological substrates. In addition, we found significant differences between zebrafish Mate7 and mammalian Mates’ substrate preferences, a finding that should be taken into consideration when using zebrafish as a model organism in toxicokinetic studies.

斑马鱼Mate7属于溶质载体蛋白超家族，尤其是多药和毒素挤出机的亚家族。它与哺乳动物的伴侣同源，在斑马鱼组织中普遍表达，在肾脏中表达最高。它已经显示出与内源性（类固醇激素）和异种化合物（药物）相互作用，这暗示着有毒化合物的外排作用。我们研究的目的是使用新开发的高通量筛选（HTS）Mate7分析方法分析环境污染物与斑马鱼Mate7的相互作用。全长斑马鱼伴侣7从源自雄性肾脏的斑马鱼cDNA中获得序列，并在基因工程改造的HEK293 Flp-In细胞中实现了Mate7的稳定表达。然后将稳定的Mate7转染子用于开发和优化新的HTS细胞摄取方案，并以DAPI和ASP +作为模型荧光底物。所开发的测定法用于鉴定斑马鱼Mate 7相互作用子并识别相互作用的类型。测试了一系列89种不同的环境污染物，包括工业化学品，农药和药物，并在上述所有组中鉴定出了高效的Mate7相互作用物。确定的某些抑制剂可能与环境有关，因为它们可能会损害Mate7的外排功能，降低鱼类对环境污染物的防御能力，或干扰尚未确定的生理底物的运输。此外，我们发现斑马鱼Mate7与哺乳动物伴侣的底物偏好之间存在显着差异，这一发现在毒物动力学研究中将斑马鱼用作模型生物时应予以考虑。